Preservation of myocardial function after adenoviral gene transfer in isolated myocardium

• Published in: American journal of physiology. Heart and circulatory physiology 2000-09, Vol.279 (3), p.H986-H991
• Main Authors: Lehnart, S. E, Janssen, P. M. L, Franz, W. M, Donahue, J. K, Lawrence, J. H, Marban, E, Prestle, J, Hasenfuss, G
• Format: Article
• Language: English
• Subjects: Adenoviridae - genetics; Adenoviridae - metabolism; Animals; Female; Gene Expression; Gene Expression Regulation, Viral; Gene Transfer Techniques; Genes, Reporter - genetics; Genetic Vectors - genetics; Genetic Vectors - pharmacology; Heart - drug effects; Heart - physiology; Heart - virology; Lac Operon - genetics; Luciferases - genetics; Myocardial Contraction - drug effects; Myocardial Contraction - genetics; Myocardium - metabolism; Organ Culture Techniques; Rabbits; Receptors, Adrenergic, beta - drug effects; Receptors, Adrenergic, beta - metabolism; Signal Transduction - drug effects; Transgenes - genetics
• ISSN: 0363-6135; 1522-1539
• DOI: 10.1152/ajpheart.2000.279.3.h986

1  Abteilung Kardiologie und Pneumologie, Universität Göttingen, D-37075 Göttingen, Germany; 2  Medizinische Klinik II, Universität zu Lübeck, D-23538 Lübeck, Germany; and 3  Section of Molecular and Cellular Cardiology, Johns Hopkins University Medical School, Baltimore, Maryland 21205 Adenoviral gene transfer to the heart represents a promising model for structure-function analyses. Rabbit hearts were subjected to an ex vivo perfusion protocol that achieves gene transfer in >90% of cardiac myocytes. Contractile function of isolated myocardial preparations of these hearts was then observed for 2 days in a recently developed trabecula culture system. In sham-infected hearts, the initial developed force (F init ) (15.6 ± 3.7 mN/mm 2 ; n  = 12) did not change significantly after 48 h (17.0 ± 1.9   mN/mm 2 ; P  = 0.46). In adenovirus-infected preparations, F init (14.3   ± 1.8 mN/mm 2 ; n  = 21) did not significantly differ from the control ( P  = 0.75) and was unchanged after 48 h (15.3 ± 2.5 mN/mm 2 ; P  = 0.93). After 2 days of continuous contractions, we observed homogenous and high-level expression of the reporter genes LacZ coding for -galactosidase and Luc coding for firefly luciferase. Luciferase activity increased more than 2,500-fold from background levels of 8.7 × 10 3 ± 5.0 × 10 3 relative light units (RLU)/mg protein (from hearts transfected with promotorless adenovirus with luciferase transgene construct AdNULLLuc, n  = 5) to 23.4 × 10 6 ± 11.1 × 10 6 RLU/mg protein (from hearts tranfected with adenovirus with Rous sarcoma virus promotor and luciferase transgene construct AdRSVLuc, n  = 5) in infected myocardial preparations ( P